metastat and Neoplasm-Metastasis

Matrix metalloproteinases (MMPs) play a significant role during the development and metastasis of prostate cancer (CaP). CaP cells secrete high levels of MMPs and low levels of endogenous MMP inhibitors (TIMPs), thus creating an excess balance of MMPs. Established CaP cell lines that express high levels of MMPs frequently metastasize to the bone and the lungs. Drugs such as Taxol and alendronate that reduce cell motility and calcium metabolism reduce bony metastasis of xenografted CaP tumors. We tested several synthetic, nontoxic inhibitors of MMPs that can be administered orally, including doxycycline (DC) and chemically modified tetracyclines (CMTs) on CaP cells in vitro and on a rat CaP model in vivo. Among several anti-MMP agents tested, CMT-3 (6-deoxy, 6-demethyl,4-de-dimethylamino tetracycline) showed highest activity against CaP cell invasion and cell proliferation. Micromolar concentration of CMT-3 and DC inhibited both the secretion and activity of MMPs by CaP cells. When tested for in vivo efficacy in the Dunning rat CaP model by daily oral gavage, CMT-3 and DC both reduced the lung metastases (> 50%). CMT-3, but not DC, inhibited tumor incidence (55 +/- 9%) and also reduced the tumor growth rate (27 +/- 9.3%). More significantly, the drugs showed minimum systemic toxicity. Ongoing studies indicate that CMT-3 may inhibit the skeletal metastases of CaP cells and delay the onset of paraplegia due to lumbar metastases. These preclinical studies provide the basis for clinical trials of CMT-3 for the treatment of metastatic disease.

Topics: Animals; Antineoplastic Agents; Bone Neoplasms; Cell Survival; Doxycycline; Humans; Lung Neoplasms; Male; Metalloendopeptidases; Neoplasm Metastasis; Prostatic Neoplasms; Protease Inhibitors; Rats; Tetracyclines; Transplantation, Heterologous; Tumor Cells, Cultured

Anti-angiogenesis is an exciting new approach to anticancer therapy. COL-3, a tetracycline derivative, is a novel anti-angiogenesis agent with potent preclinical anticancer activity. During the conduct of a phase 1 clinical trial for refractory metastatic cancer at the National Institutes of Health, we observed 3 individuals who developed phototoxicity followed by clinical and laboratory features of drug-induced lupus.. Three of 35 patients treated with COL-3 developed sunburnlike eruptions accompanied by fever and a positive antinuclear antibody titer within 8 to 29 days of starting treatment. Two of 3 had positive antihistone antibody levels and arthralgia. One patient had marked systemic manifestations including pulmonary infiltrates and elevated erythrocyte sedimentation rate remittent for more than 1 year after discontinuing COL-3 treatment. The other 2 patients' symptoms and rash abated within 2 weeks of discontinuing therapy although the serologic markers remained abnormal for the duration of follow-up.. COL-3 is the second tetracycline derivative to be implicated in the development of drug-induced lupus. A sunburnlike eruption immediately preceded or accompanied the systemic and serologic changes in these 3 patients. The rapid onset and the phototoxic appearance of the accompanying eruptions might suggest that damage to the keratinocytes caused the formation of neoantigens to which autoantibodies formed.

Topics: Administration, Topical; Aged; Anti-Inflammatory Agents; Clobetasol; Female; Follow-Up Studies; Glucocorticoids; Humans; Lupus Erythematosus, Cutaneous; Male; Matrix Metalloproteinase Inhibitors; Middle Aged; Neoplasm Metastasis; Ointments; Prednisone; Protease Inhibitors; Tetracyclines; Time Factors

Paclitaxel, a chemotherapeutic agent used in the treatment of breast cancer and other solid tumor types, including ovarian and lung, causes a dose‑dependent neuropathic pain, which limits its use. Chemically modified tetracycline‑3 (COL‑3) has anticancer properties and was previously reported to inhibit neuroinflammation and protect against paclitaxel‑induced neuropathic pain (PINP) in mice models. However, it is not known whether it affects the anticancer activities of paclitaxel. Thus, the aim of the present study was to evaluate the effect of COL‑3 on the anticancer activity of paclitaxel on the breast cancer cell lines MCF‑7 (estrogen receptor‑positive), pII [estrogen receptor‑negative (ER‑ve)] and MDA‑MB‑231 (ER‑ve). Cell proliferation, apoptosis and cell cycle stage were determined using an MTT assay, Annexin V/7‑aminoactinomycin D and flow cytometry. The expression of various signaling molecules was determined with ELISA‑based proteome profiling and western blotting. Additionally, the degree of cell invasion was determined with a Matrigel assay and caspase‑3 activity was determined with a colorimetric assay. Treatment with paclitaxel or COL‑3 alone inhibited cell proliferation in a concentration‑dependent manner in all cell lines. The anti‑proliferative effects of paclitaxel and COL‑3 in combination varied from synergism against MDA‑MB‑231 and pII cells to notably additive and slight antagonism against MCF‑7 cells. In the highly proliferative and invasive pII cells, the observed synergistic anti‑proliferative effect was partially through the induction of apoptosis via modulation of caspase‑3 levels and activity, and P70S6K phosphorylation, but not cell cycle arrest. COL‑3 inhibited the invasion of pII cells in a concentration‑dependent manner partially through inhibiting total matrix metalloproteinase activity. The combination regimen significantly inhibited the expression of two proteases, ADAM metallopeptidase with thrombospondin type 1 motif 1 and proteinase 3. In conclusion, the combination of paclitaxel and COL‑3 indicated additive to synergistic anti‑proliferative effects on breast cancer cells mediated partially via the induction of apoptosis. The combination regimen could further inhibit invasion and metastasis. Thus, COL‑3 could be a beneficial adjunct to a paclitaxel‑based anticancer regimen to improve therapeutic outcome and reduce the adverse effects of paclitaxel, primarily PINP.

Topics: Apoptosis; Breast Neoplasms; Caspase 3; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Drug Synergism; Female; Humans; MCF-7 Cells; Neoplasm Invasiveness; Neoplasm Metastasis; Paclitaxel; Tetracyclines

COL-3 is an oral, lipophilic, tetracycline analog that has been administered to patients with metastatic cancer. Preliminary assessment of COL-3 in 35 patients with refractory metastatic carcinoma demonstrated apparent nonlinear pharmacokinetics with highly variable oral clearance (63.9% coefficient of variance [CV]). To elucidate possible sources of variability of COL-3 pharmacokinetics in vivo, in vitro plasma protein binding and in vitro metabolism were explored along with in vivo pharmacokinetics using compartmental modeling. The variability in the overall clearance and urinary excretion of COL-3 was also assessed. COL-3 had a long terminal half-life (median = 59.8 h), large apparent volume of distribution (median = 50.2 L), and low apparent clearance (median = 9.93 mL/min). Only adjusted ideal body weight decreased the variability in total apparent clearance. There was nonsaturable plasma protein binding of COL-3 (fu = 5.5%), with the majority of binding to albumin. The renal route of elimination is negligible, with 0.06% of unchanged COL-3 and 3.31% COL-3 glucuronide excreted in the first 6 days. COL-3 is not metabolized by phase I metabolism but does undergo glucuronidation in vitro by UGT1A1, UGT1A3, UGT1A9, and UGT2B7 and in vivo, as evidenced by COL-3 glucuronides in the urine (median = 13.6% of the total dose). COL-3 exhibits nonlinear pharmacokinetics, possibly due to dissolution rate-limited absorption.

Topics: Administration, Oral; Adult; Aged; Cytochrome P-450 Enzyme System; Dose-Response Relationship, Drug; Female; Glucuronides; Humans; Male; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases; Middle Aged; Neoplasm Metastasis; Orosomucoid; Pharmacokinetics; Protein Binding; Serum Albumin; Tetracyclines; Time Factors; Treatment Failure

Topics: Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Apoptosis; Cell Division; Doxycycline; Drug Interactions; Epithelial Cells; Flurbiprofen; Humans; Lung Neoplasms; Male; Metalloendopeptidases; Neoplasm Metastasis; Prostatic Neoplasms; Protease Inhibitors; Rats; Regression Analysis; Tetracyclines; Tumor Cells, Cultured